CRISPR/Cas9 Antibody 4G10 (sample size)

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Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9Western blot was performed on protein extracts from HEK293T cells transfected with Cas9 (lane 1) or from untransfected cells (lane 2) using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200216), diluted 1:5,000 in PBS-T containing 0.5% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.

Figure 2. IP using the Diagenode monoclonal antibody directed against CRISPR/Cas9IP was performed on whole cell extracts from HEK293T cells transfected with a Cas9 expression vector (lane 1, 3 and 5), or untransfected cells (lane 2 and 4) using 5 µg of the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200216, lane 3 and 4) or with an equal amount of IgG, used as a negative control (lane 5). The immunoprecipitated proteins were subsequently analysed by Western blot with the polyclonal Cas9 antibody (Cat. No. C15310258, diluted 1:5,000). Lane 1 and 2 show the result of the input.

Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9HEK293T cells were transiently transfected with a Cas9 expression vector. The cells were fixed with 4% formaldehyde, permeabilized in 0,1% Triton X-100 and blocked in PBS containing 5% BSA. The cells were stained with the Cas9 antibody diluted 1;400 at 4°C o/n, followed by incubation with an anti mouse secondary antibody coupled to AF596 for 1 h at RT (left). Nuclei were counter-stained with DAPI (middle). A merge of the two stainings is shown on the right.

When doing CRISPR experiments, it is essential to verify the expression levels of the Cas9 to ensure high-quality results. Diagenode's new CRISPR/Cas9 4G10 antibody is extremely specific, able to recognize different Cas9 fusion proteins (even those used for CRISPRi experiments) and compatible with many biochemical assays, such as immunofluorescence, western blot and immunoprecipitation. Among the different antibodies in the market, Diagenode's 4G10 CRISPR/Cas9 is our antibody of choice.

WBWestern blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.
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Guidelines for optimized gene knockout using CRISPR/Cas9Campenhout CV et al.CRISPR/Cas9 technology has evolved as the most powerful approach to generate genetic models both for fundamental and preclinical research. Despite its apparent simplicity, the outcome of a genome-editing experiment can be substantially impacted by technical parameters and biological considerations. Here, we present ...

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